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Kit for tigecycline drug sensitivity detection and method for tigecycline drug sensitivity detection by using same

A technology of tigecycline and kits, applied in biochemical equipment and methods, microbial measurement/testing, Raman scattering, etc., can solve the problems of complex operation, difficult standardization, and high MIC value, and achieve simple and accurate operation The effect of high precision and high sensitivity

Active Publication Date: 2022-01-28
QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0003] At present, there are many methods for testing tigecycline AST in clinical laboratories. However, because tigecycline is prone to oxidative degradation and unstable physical and chemical properties, there are many difficulties and operational errors in the AST of this drug, and it is not easy to standardize
For example, the tigecycline AST gold standard micro broth dilution method is complicated and time-consuming; the MIC value detected by the gradient diffusion method is high; the MTS test strip is only suitable for scientific research; some commercial systems detect tigecycline drugs Sensitivity results lack sufficient drug-resistant strains in the verification stage, and the MIC is high, false drug-resistant strains increase, and some detection system results also have high intermediary-related errors. Therefore, it is urgent to find a method in clinical diagnosis. Quickly and accurately obtain tigecycline susceptibility results

Method used

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  • Kit for tigecycline drug sensitivity detection and method for tigecycline drug sensitivity detection by using same
  • Kit for tigecycline drug sensitivity detection and method for tigecycline drug sensitivity detection by using same
  • Kit for tigecycline drug sensitivity detection and method for tigecycline drug sensitivity detection by using same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1, research on optimum heavy water concentration

[0040] 1. Experimental Preparation

[0041] 1) Culture medium preparation

[0042] Preparation of CAMHB medium: Weigh 2.1g of MUELLER-HINTON BROTH (M-H broth medium, referred to as MH medium) powder and dissolve it in 100mL ddH 2 O, add Ca 2+ solution and Mg 2+ solution, so that the final concentrations were 25mg / L and 12.5mg / L, respectively, and sterilized under high temperature and high pressure at 121°C for 15min.

[0043] 2) Preparation of bacterial suspension

[0044] The frozen standard Acinetobacter baumannii strain (Acinetobacter baumannii, ATCC19606) was inoculated into Columbia blood plates and cultured overnight in an incubator at 35°C±2°C. Pick a single colony and inoculate it into CAMHB medium, incubate at 35°C±2°C, and adjust to 0.5 McFarland turbidity.

[0045] 2. Effect of heavy water concentration on strain CD-ratio

[0046] Add the ATCC19606 bacterial solution to CAMHB medium containi...

Embodiment 2

[0055] Embodiment 2, drug susceptibility test of tigecycline

[0056] 1. Experimental Preparation

[0057] 1) Culture medium preparation

[0058] 1×CAMHB medium: Weigh 2.1g of MUELLER-HINTON BROTH powder, dissolve in 100mL ddH 2 O, add Ca 2+ solution and Mg 2+ Solution, so that the final concentration is 25mg / L, 12.5mg / L respectively, sterilized under high temperature and high pressure at 121℃ for 15min;

[0059] 2×CAMHB medium: Weigh 4.2g of MUELLER-HINTON BROTH powder, dissolve in 100mL ddH 2 O, add Ca 2+ solution and Mg 2+ solution, so that the final concentrations were 50mg / L and 25mg / L, respectively, and sterilized under high temperature and high pressure at 121°C for 15min.

[0060] 2) Drug preparation

[0061] Tigecycline (Tigecycline, Tgc) stock solution: use ddH 2 O Prepare tigecycline mother solution with a concentration of 16 mg / mL, aliquot into centrifuge tubes, wrap them in tin foil, and store at -80°C. (Note: The whole process needs to be operated and s...

Embodiment 3

[0080] Embodiment 3, tigecycline susceptibility detection kit and application

[0081] 1. Kit composition

[0082] Table 1 Composition of the kit

[0083]

[0084] 2. Kit application

[0085] 1) Sample preparation: Pick a single colony on the plate of the clinical sample to be tested (pathogenic bacteria shown in Table 2, strain source references Zhang Z, Chen M, Yu Y, et al.Antimicrobial susceptibility among gram-positive and gram- negative blood-borne pathogens collected between 2012-2016as part of the Tigecycline Evaluation and Surveillance Trial[J]. Antimicrobial Resistance&Infection Control, 2018, 7(1).) Inoculated in CAMHB medium, incubated at 35℃±2℃, adjusted to 0.5 McFarland turbidity, made of bacteria suspension.

[0086] 2) Drug sensitivity plate preparation: add culture medium to the drug sensitivity plate to fully dissolve the tigecycline powder, and avoid light during the whole process.

[0087] 3) Inoculate the suspension of the bacteria to be tested in th...

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Abstract

The invention discloses a kit for tigecycline drug sensitivity detection and a method for tigecycline drug sensitivity detection by using the same. The kit comprises a tigecycline drug sensitive plate and incubation liquid. The tigecycline drug sensitive plate comprises a drug sensitive plate body, and the plate holes of the drug sensitive plate body are coated with tigecycline powder. The tigecycline drug sensitivity detection method comprises the following steps of: inoculating a to-be-detected strain suspension into a tigecycline drug sensitivity plate, incubating, adding heavy water into the tigecycline drug sensitivity plate, continuing to incubate for a period of time, sampling at different time points, and carrying out Raman detection; calculating data in a spectrum obtained by Raman detection of the sample to obtain the minimum inhibitory concentration eMIC-MA in the drug concentration under the condition that the metabolic inhibition level of the sample is greater than or equal to 0.8 after continuous incubation for a period of time; and judging the drug sensitivity of the to-be-detected strain tigecycline according to a result judgment standard formulated by the American Food and Drug Administration and the European Drug Sensitivity Test Committee according to the numerical value of eMIC-MA. According to the method, the tigecycline drug sensitivity result can be quickly and accurately obtained.

Description

technical field [0001] The invention relates to a kit for detecting drug sensitivity to tigecycline and a method for detecting drug sensitivity to tigecycline, belonging to the field of microbial drug sensitivity detection. Background technique [0002] Tigecycline is a glycylcycline antibiotic that was approved for marketing in the United States in 2005. Tigecycline was approved and marketed in more than 50 countries and regions around the world, and was approved for marketing in China in early 2012. Currently annotated indications: (1) Complicated skin and skin and soft tissue infections; (2) Complicated intra-abdominal infections; (3) Community-acquired bacterial pneumonia. Because tigecycline has broad-spectrum and strong antibacterial activity, it has very good curative effect on common clinical multidrug-resistant bacterial infections, and it is easy to use clinically, and it does not need to adjust the dose according to the renal function damage. Therefore, this antib...

Claims

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Application Information

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IPC IPC(8): C12Q1/18G01N21/65
CPCC12Q1/18G01N21/65Y02A50/30
Inventor 朱鹏飞戴靖刘惠杰毛昱力付晓婷张丽丽籍月彤徐健
Owner QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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